A rapid, low-cost, and disposable microfluidic thread-based isotachophoresis method was developed for the purification and preconcentration of nucleic acids from biological samples, prior to their extraction and successful analysis using quantitative polymerase chain reaction (qPCR). This approach extracts and concentrates protein-free DNA from the terminating electrolyte buffer, via a continuous sampling approach, resulting in significant focussing of the extracted DNA upon a 6 cm length nylon thread. The platform was optimised using the preconcentration of a fluorescent dye, showing a 600-fold concentration capacity within <5 min. The system was then applied to the one-step extraction of lambda DNA – an E. coli bacteriophage – spiked into whole blood, exhibiting the exclusion of PCR inhibitors. The extraction efficiency from the thread material following concentration was consistent, between 94.4–113.9%. The determination of lambda DNA in whole blood was achieved within a linear range of 1.0–1 × 10⁵ fg μL⁻¹ (20–2 × 10⁶ copies per μL). This technique demonstrates great potential for the development of thread-based affordable analytical and diagnostic devices based upon DNA and RNA isolation.

Item Type:	Refereed Article
Keywords:	Thread-based isotachophoresis, DNA extraction, DNA purification, DNA, separation science
Research Division:	Chemical Sciences
Research Group:	Analytical chemistry
Research Field:	Separation science
Objective Division:	Expanding Knowledge
Objective Group:	Expanding knowledge
Objective Field:	Expanding knowledge in the chemical sciences
UTAS Author:	Chen, L (Miss Liang Chen)
UTAS Author:	Cabot, JM (Dr Joan Cabot Canyelles)
UTAS Author:	Paull, B (Professor Brett Paull)
ID Code:	151715
Year Published:	2021
Funding Support:	Australian Research Council (DP170102572)
Web of Science® Times Cited:	5
Deposited By:	Chemistry
Deposited On:	2022-08-03
Last Modified:	2022-09-07
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